Supporting Information for Schilancitrilactones A C: Three Unique Nortriterpenoids from Schisandra lancifolia Xiao Luo,, Yi-Ming Shi,, Rong-ua Luo, Shi-ong Luo, Xiao-Nian Li, Rui-Rui Wang, Sheng-ong Li, Yong-Tang Zheng, Xue Du, Wei-Lie Xiao,*, Jian-Xin Pu, and an-dong Sun*, State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, Yunnan, P. R. China, enan University of Traditional Chinese Medicine, Zhengzhou 450008, enan, P. R. China, Key Laboratory of Animal Models and uman Disease Mechanisms of Chinese Academy of Sciences & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, Yunnan, P. R. China, and Graduate University of the Chinese Academy of Sciences, Beijing 100049, P. R. China. xwl@mail.kib.ac.cn; hdsun@mail.kib.ac.cn * Corresponding author. Phone: (86) 871-5223251. Fax: (86) 871-5216343 Kunming Institute of Botany. enan University of Traditional Chinese Medicine. Kunming Institute of Zoology. Graduate University of the Chinese Academy of Sciences. 1
Contents of Supporting Information No. Contents Page No. Contents Page 1 Detailed experimental procedures and physical-chemical properties 4 18 Figure S16. MBC spectrum of schilancitrilactone B (2) 24 2 Table S1. 1 NMR assignments of schilancitrilactones A C (1 3) 8 19 Figure S17. 1 1 CSYspectrum of schilancitrilactone B (2) 25 3 Figure S1. 1 NMR spectrum of schilancitrilactone A (1) 9 20 Figure S18. RESY spectrum of schilancitrilactone B (2) 26 4 Figure S2. 13 C NMR spectrum of schilancitrilactone A (1) 10 21 Figure S19. ESIMS of schilancitrilactone B (2) 27 5 Figure S3. SQC spectrum of schilancitrilactone A (1) 11 22 Figure S20. RESIMS of schilancitrilactone B (2) 28 6 Figure S4. MBC spectrum of schilancitrilactone A (1) 12 23 Figure S21. IR spectrum of schilancitrilactone B (2) 29 7 Figure S5. 1 1 CSYspectrum of schilancitrilactone A (1) 13 24 Figure S22. UV spectrum of schilancitrilactone B (2) 30 8 Figure S6. RESY spectrum of schilancitrilactone A (1) 14 25 Figure S23. CD spectrum of schilancitrilactone B (2) 31 9 Figure S7. ESIMS of schilancitrilactone A (1) 15 26 Figure S24. RD spectrum of schilancitrilactone B (2) 32 10 Figure S8. RESIMS of schilancitrilactone A (1) 16 27 Figure S25. 1 NMR spectrum of schilancitrilactone C (3) 33 11 Figure S9. IR spectrum of schilancitrilactone A (1) 17 28 Figure S26. 13 C NMR spectrum of schilancitrilactone C (3) 34 12 Figure S10. UV spectrum of schilancitrilactone A (1) 18 29 Figure S27. SQC spectrum of schilancitrilactone C (3) 35 13 Figure S11. CD spectrum of schilancitrilactone A (1) 19 30 Figure S28. MBC spectrum of schilancitrilactone C (3) 36 14 Figure S12. RD spectrum of schilancitrilactone A (1) 20 31 Figure S29. 1 1 CSYspectrum of schilancitrilactone C (3) 37 15 Figure S13. 1 NMR spectrum of schilancitrilactone B (2) 21 32 Figure S30. RESY spectrum of schilancitrilactone C (3) 38 16 Figure S14. 13 C NMR spectrum of schilancitrilactone B (2) 22 33 Figure S31. ESIMS of schilancitrilactone C (3) 39 17 Figure S15. SQC spectrum of schilancitrilactone B (2) 23 34 Figure S32. RESIMS of schilancitrilactone C (3) 40 2
35 Figure S33. IR spectrum of schilancitrilactone C (3) 41 39 Figure S37. X-ray crystal structrue of schilancitrilactone A (1) 45 36 Figure S34. UV spectrum of schilancitrilactone C (3) 42 40 Figure S38. X-ray crystal structrue of schilancitrilactone B (2) 46 37 Figure S35. CD spectrum of schilancitrilactone C (3) 43 41 Figure S39. X-ray crystal structrue of schilancitrilactone C (3) 47 38 Figure S36. RD spectrum of schilancitrilactone C (3) 44 3
Detailed experimental procedures and physical-chemical properties 1. General Experimental procedures ptical rotations were measured with a JASC DIP 370 digital polarimeter. UV data were obtained on a Shimadzu UV 2401A spectrophotometer. CD spectra were measured on a Chirascan instrument. A BioRad FtS 135 spectrophotometer was used for scanning IR spectroscopy with KBr pellets. 1D and 2D NMR spectra were recorded on Bruker AM 400 and DRX 500 spectrometers. Unless otherwise specified, chemical shifts (δ) were expressed in ppm with reference to the solvent signals. igh resolution electrospray ionization (RESIMS) were performed on a VG Autospec 3000 spectrometer under 70 ev. X-ray diffraction was realized on a Bruker APEX DU instrument. Column chromatography was performed with silica gel (200 300 mesh; Qingdao Marine Chemical, Inc., Qingdao, People s Republic of China), Lichroprep RP-18 gel (40 63 μm, Merck, Darmstadt, Germany), or MCI gel (75 150 μm, Mitsubishi Chemical Corporation, Tokyo, Japan). Semipreparative PLC was performed on an Agilent 1200 liquid chromatography with a Zorbax SB C 18, 9.4 mm 25 cm column. Fractions were monitored by TLC and spots were visualized by heating silica gel plates sprayed with 10% 2 S 4 in Et. All solvents including petroleum ether (60 90 o C) were distilled prior to use. 2. Plant material The leaves and stems of Schisandra lancifolia were collected from Nujiang prefecture of Yunnan province, People s Republic of China, in ctober 2007. Voucher specimens (KIB 20071007) were deposited at the State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, and were identified by Prof. ong Wang. 3. Extraction, isolation and purification of compounds 1 3 The air-dried and powdered leaves and stems (5.0 kg) were extracted with 70% aqueous Me 2 C (4 6 L, 2 days each) at room temperature, and concentrated under reduced pressure to give crude extract, which was partitioned between 2 and EtAc. The EtAc part (125 g) was chromatographed on a silica gel column with a gradient elution of CCl 3 /Me 2 C (1:0 to 0:1) to furnish six fractions A F. Fraction C 4
(12.5 g) was subject to further separation over columns: first MCI eluted with 90% Me, then on RP-18 with a gradient elution of Me/ 2 (2:8 to 1:0) to yield fractions C1 C5. Subsequently fraction C4 (0.5 g) was purified by a silica gel column (petroleum ether/me 2 C 3 4:1 to 1:0) to give subfractions C4b and C4c. Subfraction C4b was then run on Sephadex L-20 eluting with methanol to afford five subfractions I-V. Subfraction C4bII subjected to purified by smipreparative PLC (3 ml/min, detector UV λ max 275 nm, Me/ 2 55:45) to yield 2 (2 mg) and 3 (8 mg). Subfraction C4c was purified by smipreparative PLC (3 ml/min, detector UV λ max 275 nm, MeCN/ 2 35:65) to yield 1 (5 mg). 4. Physical-chemical properties of compounds 1 3 Schilancitrilactone A (1), colorless needle crystals; [ ] 18.1 D +13.8 (c 0.07, Me); UV (Me) λ max (log ε): 274 (3.73) nm, 202 (3.45) nm; CD (Me) λ max (Δε): 270 ( 14.54); IR (KBr) ν max 3434, 2927, 1769, 1629, 1062 cm -1 ; positive ESIMS [M + Na] + m/z 567; positive RESIMS [M + Na] + m/z 567.2214 (calcd. C 29 36 10 Na [M + Na] +, 567.2206). Schilancitrilactone B (2), colorless chunk crystals; [ ] 18.1 D 41.0 (c 0.07, Me); UV (Me) λ max (log ε): 275 (3.99) nm; CD (Me) λ max (Δε): 268 ( 9.64); IR (KBr) ν max 3440, 2973, 2932, 1766, 1629, 1060 cm -1 ; positive ESIMS [M + Na] + m/z 509; positive RESIMS [M + Na] + m/z 509.2168 (calcd. C 27 34 8 Na [M + Na] +, 509.2151). Schilancitrilactone C (3), colorless prism crystals; [ ] 18.1 D +122.2 (c 0.07, Me); UV (Me) λ max (log ε): 275 (3.98) nm; CD (Me) λ max (Δε): 271 (+24.29); IR (KBr) ν max 3440, 2928, 2931, 1765, 1628, 1054 cm -1 ; positive ESIMS [M + Na] + m/z 509; positive RESIMS [M + Na] + m/z 509.2146 (calcd. C 27 34 8 Na [M + Na] +, 509.2151). X-ray Crystallographic Analysis of schilancitrilactone A (1): 2C 29 36 10 + 3 2, M = 1143.20, colorless needle, size 0.11 0.11 0.60 mm 3, monoclinic, space group P21; a = 20.2712(5) Å, b = 6.8588(2) Å, c = 21.9489(6) Å, α = γ = 90.00, β = 109.7560(10), V = 2872.07(13) Å 3, T = 100(2) K, Z = 2, ρ calcd. = 1.322 g/cm 3, μ(cu Kα) = 0.854 mm 1, F(000) = 1220, 22936 reflections in h( 24/23), k( 7/6), l( 26/26), measured in the range 9.77 θ 68.03, completeness θmax = 91.6%, 7831 5
independent reflections, R int = 0.0381, 7487 reflections with F 2 2σ F 2, 741 parameters, 1 restraints, GF = 1.044. Final R indices: R 1 = 0.0589, wr 2 = 0.1636. R indices (all data): R 1 = 0.0608, wr 2 = 0.1671. Flack parameter 0.12(18), largest difference peak and hole = 0.450 and 0.266 e Å 3. The intensity data for 1 were collected on a Bruker APEX DU diffractometer using graphitemonochromated Cu Kα radiation. The structure of 1 was solved by direct methods (SELXS97), expanded using difference Founier techniques, and refined by the program and full-matrix least-squares calculations. The nonhydrogen atoms were refined anisotropically, and hydrogen atoms were fixed at calculated positions. Crystallographic data for the structure of 1 have been deposited in the Cambridge Crystallographic Data Centre (deposition number CCDC 861514). Copies of the data can be obtained free of charge from the CCDC via www.ccdc.cam.ac.uk. X-ray Crystallographic Analysis of schilancitrilactone B (2): C 27 34 8, M = 486.54, colorless chunk, size 0.18 0.35 0.40 mm 3, monoclinic, space group P21; a = 10.40640(10) Å, b = 6.95520(10) Å, c = 18.3789(2) Å, α = γ = 90.00, β = 102.0530(10), V = 1300.91(3) Å 3, T = 296(2) K, Z = 2, ρ calcd. = 1.242 g/cm 3, μ(cu Kα) = 0.750 mm 1, F(000) = 520, 8415 reflections in h( 12/10), k( 8/8), l( 21/21), measured in the range 2.46 θ 66.55, completeness θmax = 94.9%, 3723 independent reflections, R int = 0.0369, 3680 reflections with F 2 2σ F 2, 323 parameters, 1 restraints, GF = 1.057. Final R indices: R 1 = 0.0551, wr 2 = 0.1617. R indices (all data): R 1 = 0.0554, wr 2 = 0.1625. Flack parameter 0.00(19), largest difference peak and hole = 0.296 and 0.382 e Å 3. The intensity data for 2 were collected on a Bruker APEX DU diffractometer using graphitemonochromated Cu Kα radiation. The structure of 2 was solved by direct methods (SELXS97), expanded using difference Founier techniques, and refined by the program and full-matrix least-squares calculations. The nonhydrogen atoms were refined anisotropically, and hydrogen atoms were fixed at calculated positions. Crystallographic data for the structure of 2 have been deposited in the Cambridge Crystallographic Data Centre (deposition number CCDC 861515). Copies of the data can be obtained free of charge from the CCDC via www.ccdc.cam.ac.uk. X-ray Crystallographic Analysis of schilancitrilactone C (3): C 27 34 8 + 2, M = 504.56, colorless prism, size 0.54 0.80 0.80 mm 3, orthorhombic, space group 6
P212121; a = 10.2430(4) Å, b = 10.5955(5) Å, c = 23.2804(9) Å, α = β = γ = 90.00, V = 2526.62(18) Å 3, T = 100(2) K, Z = 4, ρ calcd. = 1.326 g/cm 3, μ(cu Kα) = 0.821 mm 1, F(000) = 1080, 10991 reflections in h( 11/12), k( 10/11), l( 27/27), measured in the range 3.80 θ 69.47, completeness θmax = 92.9%, 4261 independent reflections, R int = 0.0326, 4254 reflections with F 2 2σ F 2, 334 parameters, 0 restraint, GF = 1.103. Final R indices: R 1 = 0.0341, wr 2 = 0.0872. R indices (all data): R 1 = 0.0341, wr 2 = 0.0872. Flack parameter 0.03(14), largest difference peak and hole = 0.239 and 0.261 e Å 3. The intensity data for 3 were collected on a Bruker APEX DU diffractometer using graphitemonochromated Cu Kα radiation. The structure of 3 was solved by direct methods (SELXS97), expanded using difference Founier techniques, and refined by the program and full-matrix least-squares calculations. The nonhydrogen atoms were refined anisotropically, and hydrogen atoms were fixed at calculated positions. Crystallographic data for the structure of 3 have been deposited in the Cambridge Crystallographic Data Centre (deposition number CCDC 861516). Copies of the data can be obtained free of charge from the CCDC via www.ccdc.cam.ac.uk. 7
Table S1 1 NMR Data for Compounds 1 3 (CDCl 3, in ppm) a position 1 2 3 (mult., J in z) (mult., J in z) (mult., J in z) 1 4.19 br d (5.0) 2 2.69 d (18.1) 2 2.76 br dd (18.1, 5.0) 5 2.77 m 2.47 m 2.47 m 6 1.52 m 1.67 overlap 1.65 overlap 6 1.40 m 1.50 m 1.48 m 7 1.86 overlap 1.67 overlap 1.66 overlap 7 1.86 overlap 1.67 overlap 1.66 overlap 8 2.38 m 2.23 m 2.18 m 10 3.38 m 3.37 m 11 2.05 m 2.06 overlap 2.02 overlap 11 1.70 m 1.86 dd (13.5, 5.0) 1.84 dd (13.6, 5.0) 12 2.43 m 2.51 m 2.50 m 14 4.47 dd (7.9, 5.7) 4.41 dd (7.2, 4.2) 4.37 dd (7.3, 4.3) 16 2.44 ABd (17.7) 2.45 ABd (18.0) 2.63 ABd (18.0) 16 2.63 ABd (17.7) 2.67 ABd (18.0) 2.72 ABd (18.0) 19 1.91 overlap 2.10 m 2.03 overlap 19 1.91 overlap 1.74 overlap 1.73 ABd (14.4) 20 3.34 m 3.34 m 3.37 m 21 1.25 d (6.7) 1.26 d (6.8) 1.20 d (6.8) 22 4.97 d (10.5) 4.99 d (10.5) 5.28 d (9.8) 24 6.98 br d (0.8) 6.99 br d (1.1) 7.02 br d (1.0) 27 2.01 s 2.02 s 2.00 s 28 1.19 s 1.20 s 1.20 s 29 3.57 d (12.0) 1.32 s 1.31 s 3.40 d (12.0) 30 1.06 s 1.44 s 1.43 s a Data for compound 1 3 were recorded at 500 Mz, and the assignments were base on DEPT, SQC, MBC, CSY, and RESY experiments. 8
1 Figure S1. 1 NMR spectrum of schilancitrilactone A (1) 9
1 Figure S2. 13 C NMR spectrum of schilancitrilactone A (1) 10
1 Figure S3. SQC spectrum of schilancitrilactone A (1) 11
1 Figure S4. MBC spectrum of schilancitrilactone A (1) 12
1 Figure S5. 1 1 CSY spectrum of schilancitrilactone A (1) 13
1 Figure S6. RESY spectrum of schilancitrilactone A (1) 14
1 Figure S7. ESIMS of schilancitrilactone A (1) 15
1 Figure S8. RESIMS of schilancitrilactone A (1) 16
1 Figure S9. IR spectrum of schilancitrilactone A (1) 17
1 Figure S10. UV spectrum of schilancitrilactone A (1) 18
1 Figure S11. CD spectrum of schilancitrilactone A (1) 19
1 Figure S12. RD spectrum of schilancitrilactone A (1) 20
2 Figure S13. 1 NMR spectrum of schilancitrilactone B (2) 21
2 Figure S14. 13 C NMR spectrum of schilancitrilactone B (2) 22
2 Figure S15. SQC spectrum of schilancitrilactone B (2) 23
2 Figure S16. MBC spectrum of schilancitrilactone B (2) 24
2 Figure S17. 1 1 CSY spectrum of schilancitrilactone B (2) 25
2 Figure S18. RESY spectrum of schilancitrilactone B (2) 26
2 Figure S19. ESIMS of schilancitrilactone B (2) 27
2 Figure S20. RESIMS of schilancitrilactone B (2) 28
2 Figure S21. IR spectrum of schilancitrilactone B (2) 29
2 Figure S22. UV spectrum of schilancitrilactone B (2) 30
2 Figure S23. CD spectrum of schilancitrilactone B (2) 31
Figure S24. RD spectrum of schilancitrilactone B (2) 2 32
3 Figure S25. 1 NMR spectrum of schilancitrilactone C (3) 33
3 Figure S26. 13 C NMR spectrum of schilancitrilactone C (3) 34
3 Figure S27. SQC spectrum of schilancitrilactone C (3) 35
3 Figure S28. MBC spectrum of schilancitrilactone C (3) 36
3 Figure S29. 1 1 CSY spectrum of schilancitrilactone C (3) 37
3 Figure S30. RESY spectrum of schilancitrilactone C (3) 38
3 Figure S31. ESIMS of schilancitrilactone C (3) 39
3 Figure S32. RESIMS of schilancitrilactone C (3) 40
3 Figure S33. IR spectrum of schilancitrilactone C (3) 41
3 Figure S34. UV spectrum of schilancitrilactone C (3) 42
3 Figure S35. CD spectrum of schilancitrilactone C (3) 43
Figure S36. RD spectrum of schilancitrilactone C (3) 3 44
1 Figure S37. X-ray crystal structrue of schilancitrilactone A (1) 45
2 Figure S38. X-ray crystal structrue of schilancitrilactone B (2) 46
3 Figure S39. X-ray crystal structrue of schilancitrilactone C (3) 47